High Sensitive Acetylserotonin O-methyltransferase ELISA Test Kit Human ASMT Sandwich ELISA Kit

Cat.No E6644Hu

Intended Use

This sandwich kit is for the accurate quantitative detection of Human Acetylserotonin O-methyltransferase (also known as ASMT) in serum, plasma, cell culture supernates, cell lysates, tissue homogenates.

Assay Principle

This ELISA Test Kit is an Enzyme-Linked Immunosorbent Assay (ELISA). The plate has been pre-coated with Human ASMT antibody. ASMT present in the sample is added and binds to antibodies coated on the wells. And then biotinylated Human ASMT Antibody is added and binds to ASMT in the sample. Then Streptavidin-HRP is added and binds to the Biotinylated ASMT antibody. After incubation unbound Streptavidin-HRP is washed away during a washing step. Substrate solution is then added and color develops in proportion to the amount of Human ASMT. The reaction is terminated by addition of acidic stop solution and absorbance is measured at 450 nm.

Standard Curve Range: 0.05ng/ml - 20ng/ml

Sensitivity: 0.016ng/ml

Size: 96 wells

Storage: Store the reagents at 2-8°C. For over 6-month storage refer to the expiration date keep it at -20°C. Avoid repeated thaw cycles. If individual reagents are opened it is recommended that the kit be used within 1 month.

*This product is for research use only, not for use in diagnosis procedures. It’s highly recommend to read this instruction entirely before use.

Reagent Provided

Material Required But Not Supplied

• 37°C±0.5°C incubator
• Absorbent paper
• Precision pipettes and disposable pipette tips
• Clean tubes
• Deionized or distilled water
• Microplate reader with 450 ± 10nm wavelength filter

Assay Procedure

1. Prepare all reagents, standard solutions and samples as instructed. Bring all reagents to room temperature before use. The assay is performed at room temperature.

2. Determine the number of strips required for the assay. Insert the strips in the frames for use. The unused strips should be stored at 2-8°C.

3. Add 50μl standard to standard well. Note: Don’t add antibody to standard well because the standard solution contains biotinylated antibody.

4. Add 40μl sample to sample wells and then add 10μl anti-ASMT antibody to sample wells, then add 50μl streptavidin-HRP to sample wells and standard wells ( Not blank control well ). Mix well. Cover the plate with a sealer. Incubate 60 minutes at 37°C.

5. Remove the sealer and wash the plate 5 times with wash buffer. Soak wells with at least 0.35 ml wash buffer for 30 seconds to 1 minute for each wash. For automated washing, aspirate all wells and wash 5 times with wash buffer, overfilling wells with wash buffer. Blot the plate onto paper towels or other absorbent material.

6. Add 50μl substrate solution A to each well and then add 50μl substrate solution B to each well. Incubate plate covered with a new sealer for 10 minutes at 37°C in the dark.

7. Add 50μl Stop Solution to each well, the blue color will change into yellow immediately.

8. Determine the optical density (OD value) of each well immediately using a microplate reader set to 450 nm within 10 minuets after adding the stop solution.

Summary

1. Prepare all reagents, samples and standards.

2. Add sample and ELISA reagent into each well. Incubate for 1 hour at 37°C.

3. Wash the plate 5 times.

4. Add substrate solution A and B. Incubate for 10 minutes at 37°C.

5. Add stop solution and color develops.

6. Read the OD value within 10 minutes.